新型固相竞争性时间分辨荧光免疫检测血清孕酮方法的建立

A Novel Solid—phase Competitive TRFIA for Serum Progesterone

本研究以血清孕酮为对象,在国内首次建立了小分子类固醇激素的时间分辨荧光免疫分析法。将半抗原——载体蛋白分子即孕酮——BSA 包被微量滴定条后,再行无水乙醇处理,制成固相抗原;纯化的抗孕酮抗体 IgG 与生物素偶联;E_u^3+则与链亲和素标记,作为通用标记物。该法曲线范围是0.1～100ng/ml,灵敏度为0.05ng/ml,平均批内及批间 C、V 分别为6.45%及8.93%,平均回收为89.95%,血清线性稀释实验测得值与期望值接近.40例样本同时用 TRFIA 及 RIA 测量,相关性良好,r=0.9619。

The acticle describes a time—resolved fluoroimmunoassay for serum progesterone,based on a competitive solid—phase antigen technique。Progesterone—BSA conjugates are coated onto the white opaque microtitration wells。Anti—progesterone antibody is biotinylated.Eu^(3+) are labelled to streptavidin as the tracer.The assay standard curve covers a range of 0.1—100ng/ml.The lowest detecting limit is 0.05ng/ml。The intra and inter C·V s are 6.45% and 8.93% respec- tively.The average recovery is 89.5%.40woman samples are measured with RIA and present TRFIA and the results are satisfactory,giving a correlation coefficient of 0.9619.The perfor- mance charicter—istics of the assay are discussed and compared with other known immunoas- says.